吸水链霉菌应城变种质粒的分离研究

ISOLATION AND STUDIES OF THE PLASMID FROM STREPTOMYCES HYGROSCOPICUS VAR. YINGCHENGENSIS

采用蔗糖浓度为13％的YEME培养基培养吸水链霉菌应城变种10-22菌株,能获得生长量较高且分散状况良好的菌丝体。在探索链霉菌质粒DNA的分离方法时,以T.Kieser的方法为基础,提高国产溶菌酶的用量和延长酶解时间,首次从吸水链霉菌应城变种中分离到一种质粒DNA分子,将它命名为pSH_(47)。同时,通过pock形成能力的检测。进一步证实这个质粒的存在。吸水链霉菌应城变种10-22菌株经吖啶橙处理后获得的部分或全部丧失5102抗生素生物合成能力的几个阻遏突变株的质粒检测结果表明,它们都有与10-22菌株相同的质粒带。

Based on the results of the experiments, YEME medium containing 13% sucrose was suitable for the growth of antibiotic 5102 producing strain,StrePtom-yces hygroscopicus var. Yingchengensis 10-22, in which mycelia of more biomass and better dispersed state were grown.A kind of plasmid DNA, named pSH47, was first isolated fom Streptomyces hygroscopicus var. Yingchengensis 10-22 with the method improved by raising the concentration of China-made lysozyme and prolonging the ly-sozyme treatment on the basis of T. Kieser's method.The existence of pasmid pSH47 was further confirmed by testing the ability of it to confer pock formation. The same plasmid bands appeared on an agarose gel by screening for the crude extcacis from Streptomyces hygroscopicus var.Yingchengensis 10-22 and several strain 10-22's block mutants, which had been gained by treating strain 10-22 with acridine orange and lost the whole or parts of the abilities producing three kinds of antibiotics 5102.